Preparation and Identification of Anti-Human B7-H1 Monoclonal Antibody

Objective: Preparation of anti-human B7-H1 monoclonal antibody with hybridoma technique and preliminary analysis of the application. Methods: The immunized mouse spleen cells and mouse myeloma cells were fused by routine method, indirect ELISA were used to screen the hybridoma. The positive clones were subcloned acquire hybridoma cells secreting anti-human B7-H1 monoclonal antibody. The amplified hybridoma cells were injected into abdominal cavity of mouse and the ascites were collected. The antibodies in the ascites were purified and titrated, and were indentified by immunohistochemical stain of lung cancer tissues. Results: One hybridoma cell line secreting anti-human B7-H1 monoclonal antibody was acquired. The subclass of the antibody was IgG1, the titer was 1×108, the concentration was 6.76 g / L after purification, and the antibody could combine with the B7-H1 which was on the surface of lung cancer tissues specifically. Conclusion: The anti-human B7-H1 monoclonal antibody was prepared successfully, which lay a foundation for later establishment of B7-H1 detecting kit